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热带爪蟾卵母细胞作为研究细胞内钙离子信号传导的优势模型系统。

Xenopus tropicalis oocytes as an advantageous model system for the study of intracellular Ca(2+) signalling.

作者信息

Marchant J S, Parker I

机构信息

Laboratory for Cellular and Molecular Neurobiology, Department of Neurobiology & Behavior, University of California at Irvine, 92697-4550, USA.

出版信息

Br J Pharmacol. 2001 Apr;132(7):1396-410. doi: 10.1038/sj.bjp.0703922.

Abstract
  1. The purpose of this study was to compare oocytes from the pipid frogs Xenopus tropicalis and Xenopus laevis, with respect to their utility for studying Ca(2+) signalling mechanisms and for expression of heterologous proteins. 2. We show that X. tropicalis oocytes possess an intracellular Ca(2+) store that is mobilized by inositol (1,4,5) trisphosphate (IP(3)). Ca(2+) signalling is activated by endogenous lysophosphatidic acid receptors and cytosolic Ca(2+) activates a plasma membrane chloride conductance. The spatiotemporal organization of cytosolic Ca(2+) signals, from the microscopic architecture of elementary Ca(2+) 'puffs' to the macroscopic patterns of Ca(2+) spiking are closely similar to the local and global patterns of Ca(2+) release previously characterized in oocytes from X. laevis. 3. By injecting X. tropicalis oocytes with cDNA encoding an ER-targeted fluorescent protein construct, we demonstrate the capacity of the X. tropicalis oocyte to readily express heterologous proteins. The organization of ER is polarized across the oocyte, with the IP(3)-releaseable store targeted within an approximately 8 microm wide band that circumscribes the cell. 4. We conclude that the X. tropicalis oocyte shares many of the characteristics that have made oocytes of X. laevis a favoured system for studying Ca(2+) signalling mechanisms. Moreover, X. tropicalis oocytes display further practical advantages in terms of imaging depth, Ca(2+) signal magnitude and electrical properties. These further enhance the appeal of X. tropicalis as an experimental system, in addition to its greater amenability to transgenic approaches.
摘要
  1. 本研究的目的是比较滑爪蟾属的热带爪蟾和非洲爪蟾的卵母细胞,就其在研究Ca(2+)信号传导机制和异源蛋白表达方面的实用性而言。2. 我们发现热带爪蟾卵母细胞拥有一个细胞内Ca(2+)储存库,可被肌醇(1,4,5)三磷酸(IP(3))动员。Ca(2+)信号由内源性溶血磷脂酸受体激活,胞质Ca(2+)激活质膜氯电导。从基本Ca(2+)“钙瞬变”的微观结构到Ca(2+)尖峰的宏观模式,胞质Ca(2+)信号的时空组织与先前在非洲爪蟾卵母细胞中表征的Ca(2+)释放的局部和全局模式非常相似。3. 通过向热带爪蟾卵母细胞注射编码内质网靶向荧光蛋白构建体的cDNA,我们证明了热带爪蟾卵母细胞易于表达异源蛋白的能力。内质网在卵母细胞中呈极化分布,IP(3)可释放的储存库定位在围绕细胞的约8微米宽的带内。4. 我们得出结论,热带爪蟾卵母细胞具有许多使非洲爪蟾卵母细胞成为研究Ca(2+)信号传导机制的首选系统的特征。此外,热带爪蟾卵母细胞在成像深度、Ca(2+)信号强度和电特性方面显示出进一步的实际优势。除了其对转基因方法的更大适应性外,这些进一步增强了热带爪蟾作为实验系统的吸引力。

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The Xenopus oocyte: a single-cell model for studying Ca2+ signaling.非洲爪蟾卵母细胞:用于研究钙离子信号传导的单细胞模型。
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本文引用的文献

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Fire-diffuse-fire model of dynamics of intracellular calcium waves.细胞内钙波动力学的“火-扩散-火”模型
Proc Natl Acad Sci U S A. 1999 May 25;96(11):6060-3. doi: 10.1073/pnas.96.11.6060.
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Frog genetics: Xenopus tropicalis jumps into the future.青蛙遗传学:热带爪蟾迈向未来。
Trends Genet. 1998 Jul;14(7):253-5. doi: 10.1016/s0168-9525(98)01506-6.

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